Amikacin
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D-Streptamine,O-3-amino-3-deoxy-a-D-glucopyranosyl-(1®6)-O-[6-amino-6-deoxy-a-D-glucopyranosyl(1®4)]-N1-(4-amino-2-hydroxy-1-oxobutyl)-2-deoxy-,(S)-.
O-3-Amino-3-deoxy-a-D-glucopyranosyl(1®4)-O-[6-amino-6-deoxy-a-D-glucopyranosyl(1®6)]-N3-(4-amino-L-2-hydroxybutyryl)-2-deoxy-L-streptamine [37517-28-5].
»Amikacin has a potency of not less than 900µg of C22H43N5O13per mg,calculated on the anhydrous basis.
Packaging and storage—
Preserve in tight containers.
Identification—
A:
Thin-Layer Chromatographic Identification Test á201ñ—
Test solution:
6mg per mL,in water.Apply 3µL.
Standard solution:
6mg per mL,in water.Apply 3µL.
Mixed solution:
a mixture of the Test solutionand the Standard solution(1:1).Apply 3µL.
Developing solvent system:
a mixture of methanol,ammonium hydroxide,and chloroform (60:35:25).
Spray reagent:
a 1in 100solution of ninhydrin in a mixture of butyl alcohol and pyridine (100:1).
Procedure—
Proceed as directed in the chapter,except to develop the chromatogram by continuous flow for 5.5hours.Remove the plate from the chamber,allow the solvent to evaporate,and heat the plate at 110
for 15minutes.Spray the plate with Spray reagent,and immediately locate the spots:amikacin appears as a pink spot,and the spots obtained from the Test solutionand the Mixed solutioncorrespond in distance from the origin to that obtained from the Standard solution.
for 15minutes.Spray the plate with Spray reagent,and immediately locate the spots:amikacin appears as a pink spot,and the spots obtained from the Test solutionand the Mixed solutioncorrespond in distance from the origin to that obtained from the Standard solution.
B:
The retention time of the peak for amikacin in the chromatogram of the Assay preparationcorresponds to that in the chromatogram of the Standard preparation,as obtained in the Assay.
Specific rotation á781Sñ:
between +97and +105.
and +105.
Test solution:
20mg per mL,in water.
Crystallinity á695ñ:
meets the requirements.
pHá791ñ:
between 9.5and 11.5,in a solution containing 10mg per mL.
Water,Method Iá921ñ:
not more than 8.5%.
Residue on ignition á281ñ:
not more than 1.0%,the charred residue being moistened with 2mLof nitric acid and 5drops of sulfuric acid.
Assay—
Mobile phase—
Use 0.115Nsodium hydroxide.Make adjustments if necessary (see System Suitabilityunder Chromatography á621ñ).
System suitability solution—
Prepare a solution in water containing about 0.02mg of USP Amikacin RSper mLand 0.008mg of USP Kanamycin Sulfate RSper mL.
Standard preparation—
Quantitatively dissolve an accurately weighed quantity of USP Amikacin RSin water to obtain a solution having a known concentration of about 0.02mg per mL.
Assay preparation—
Transfer about 50mg of Amikacin,accurately weighed,to a 250-mLvolumetric flask,dilute with water to volume,and mix.Transfer 10.0mLof this solution to a 100-mLvolumetric flask,dilute with water to volume,and mix.
Chromatographic system (see Chromatography á621ñ)—
The liquid chromatograph is equipped with an electrochemical detector,a gold working electrode,and a pHsilver-silver chloride reference electrode,a guard column that contains packing L47,and a 4-mm ×25-cm analytical column that contains packing L47.The electrochemical detector is used in the integrated amperometric mode with a range of 300nC,an output of 1Vfull scale,a rise time of 0.5second,positive polarity,potential E=0.04V;t1=200ms;E2=0.8V;t2=190ms;E3=-0.8V;t3=190ms.The flow rate is about 0.5mLper minute.Chromatograph the System suitability solution,and measure the peak areas as directed for Procedure:the relative retention times are about 0.8for kanamycin and 1.0for amikacin;and the resolution,R,between kanamycin and amikacin is not less than 3.Chromatograph the Standard preparation,and measure the peak areas as directed for Procedure:the tailing factor is not more than 2;and the relative standard deviation for replicate injections is not more than 3%.
Procedure—
Separately inject equal volumes (about 20µL)of the Standard preparationand the Assay preparationinto the chromatograph,record the chromatograms,and measure the areas for the major peaks.Calculate the quantity,in µg,of C22H43N5O13in each mg of Amikacin taken by the formula:
2500(CE/W)(rU/rS),
in which Cis the concentration,in mg per mL,of USP Amikacin RSin the Standard preparation;Eis the designated amikacin content,in µg per mg,of USP Amikacin RS;Wis the weight,in mg,of Amikacin taken to prepare the Assay preparation;and rUand rSare the amikacin peak areas obtained from the Assay preparationand the Standard preparation,respectively.
Auxiliary Information—
Staff Liaison:William W.Wright,Ph.D.,Scientific Fellow
Expert Committee:(PA7)Pharmaceutical Analysis 7
USP28–NF23Page 116
Pharmacopeial Forum:Volume No.28(4)Page 1065
Phone Number:1-301-816-8335